RESUMO
AIMS: To investigate the anti-inflammatory activity of an invasive and Hp65-producing strain Lactococcus lactis NCDO2118 FnBPA+ (pXYCYT:Hsp65) in acute 2,4,6-trinitrobenzene sulphonic acid (TNBS)-induced colitis in mice as an innovative therapeutic strategy against Crohn's disease (CD). METHODS AND RESULTS: The pXYCYT:Hsp65 plasmid was transformed into the L. lactis NCDO2118 FnBPA+ strain, resulting in the L. lactis NCDO2118 FnBPA+ (pXYCYT:Hsp65) strain. Then, the functionality of the strain was evaluated in vitro for Hsp65 production by Western blotting and for invasion into Caco-2 cells. The results demonstrated that the strain was able to produce Hsp65 and efficiently invade eukaryotic cells. Subsequently, in vivo, the anti-inflammatory capacity of the recombinant strain was evaluated in colitis induced with TNBS in BALB/c mice. Oral administration of the recombinant strain was able to attenuated the severity of colitis by mainly reducing IL-12 and IL-17 levels and increasing IL-10 and secretory immunoglobulin A levels. CONCLUSIONS: The L. lactis NCDO2118 FnBPA+ (pXYCYT:Hsp65) strain contributed to a reduction in inflammatory damage in experimental CD. SIGNIFICANCE AND IMPACT OF THE STUDY: This study, which used L. lactis for the production and delivery of Hsp65, has scientific relevance because it shows the efficacy of this new strategy based on therapeutic protein delivery into mammalian enterocytes.
Assuntos
Proteínas de Bactérias/metabolismo , Chaperonina 60/metabolismo , Colite/terapia , Imunoglobulina A Secretora/metabolismo , Interleucina-10/metabolismo , Lactococcus lactis/fisiologia , Administração Oral , Animais , Proteínas de Bactérias/genética , Células CACO-2 , Chaperonina 60/genética , Colite/induzido quimicamente , Colite/imunologia , Sistemas de Liberação de Medicamentos , Feminino , Humanos , Inflamação/terapia , Lactococcus lactis/genética , Lactococcus lactis/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Ácido Trinitrobenzenossulfônico/toxicidadeRESUMO
AIMS: A regimen utilizing Bacille Calmette-Guerin (BCG) and another vaccine system as a booster may represent a promising strategy for the development of an efficient tuberculosis vaccine for adults. In a previous work, we confirmed the ability of Lactococcus lactis fibronectin-binding protein A (FnBPA+) (pValac:ESAT-6), a live mucosal DNA vaccine, to produce a specific immune response in mice after oral immunization. In this study, we examined the immunogenicity of this strain as a booster for the BCG vaccine in mice. METHODS AND RESULTS: After immunization, cytokine and immunoglobulin profiles were measured. The BCG prime L. lactis FnBPA+ (pValac:ESAT-6) boost group was the most responsive group, with a significant increase in splenic pro-inflammatory cytokines IL-17, IFN-γ, IL-6 and TNF-α compared with the negative control. CONCLUSIONS: Based on the results obtained here, we demonstrated that L. lactis FnBPA+ (pValac:ESAT-6) was able to increase the BCG vaccine general immune response. SIGNIFICANCE AND IMPACT OF THE STUDY: This work is of great scientific and social importance because it represents the first step towards the development of a booster to the BCG vaccine using L. lactis as a DNA delivery system.
Assuntos
Antígenos de Bactérias/imunologia , Vacina BCG/imunologia , Proteínas de Bactérias/imunologia , Citocinas/sangue , Interleucina-17/metabolismo , Lactococcus lactis/genética , Vacinas de DNA/imunologia , Animais , Antígenos de Bactérias/genética , Proteínas de Bactérias/genética , Feminino , Interleucina-6 , Lactococcus lactis/metabolismo , Camundongos , Fator de Necrose Tumoral alfaRESUMO
This paper describes the development of a technique to make a fixed bridge for a patient with severe reduction of periodontal support. Initially, the bridge was temporarily cemented over copings. However, when the super-structure was periodically removed for inspection, the copings were easily dislodged, with a risk of fracture of the abutment teeth. Consequently, the technique was modified by using lateral screws to fix the bridge to the copings. This design allows easy periodical removal, cleaning, repair and inspection of the abutment teeth.